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Details for Patent: 6,455,282

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Details for Patent: 6,455,282

Title: Cells, vectors and methods for producing biologically active TSH
Abstract:Biologically active heteropolymeric protein composed of a plurality of subunits, both subunits being synthesized in a single cell having an expression vector comprising heterologous DNA encoding the subunits. Preferably the protein is similar to the human or ungulate fertility hormones, LH and FSH.
Inventor(s): Beck; Anton (Wellsley, MA), Bernstine; Edward (Boston, MA), Hsiung; Nancy (Wellesley, MA), Reddy; Vermuni B. (Framingham, MA)
Assignee: Genzyme Corporation (Cambridge, MA)
Filing Date:Nov 02, 1992
Application Number:07/970,227
Claims:1. An expression vector encoding two subunits of thyroid stimulating hormone (TSH), each under the control of a separate promoter.

2. The expression vector of claim 1, said expression vector comprising a plasmid.

3. The expression vector of claim 1, said expression vector comprising a replicating virus.

4. A mammalian cell consisting essentially of a cell transfected by a first expression vector, said transfected cell being capable of producing a biologically active heterodimeric thyroid stimulating hormone (TSH) having first and second subunits, each of said subunits of said protein being encoded by said first expression vector under the control of a separate promoter, or progeny of said transfected cell containing the promoters and DNA sequences for said subunits imparted by said vector.

5. The cell of claim 4 wherein, in said transfected, said expression vector is autonomously replicating.

6. The cell of claim 4, wherein said transfected cell is a monkey cell.

7. The cell of claim 4, wherein said transfected cell is a mouse cell.

8. A mammalian cell consisting essentially of a cell transfected by a first expression vector and a distinct second expression vector, said transfected cell being capable of producing a biologically active heterodimeric thyroid stimulating hormone (TSH) having first and second subunits, the first subunit of said protein being encoded by said first expression vector and the second subunit of said protein being encoded by said second expression vector, or progeny of said transfected cell containing the promoters and DNA sequences for said subunits imparted by said vectors.

9. The cell of claim 8 wherein, in said transfected cell, said expression vectors are autonomously replicating.

10. The cell of claim 4 or claim 8, wherein, in said transfected cell, said first vector is a plasmid.

11. The cell of claim 4 or claim 8, wherein, in said transfected cell, transcription of each of the two different subunits of said heterodimer is under the control of an SV40 late promoter.

12. The cell of claim 4 or claim 8, wherein, in said transfected cell, transcription of one subunit of said heterodimer is under the control of the SV40 early promoter, and transcription of the other subunit of said heterodimer is under the control of the mouse metallothionein promoter.

13. The cell of claim 12, wherein, in said transfected cell, said first vector comprises at least the 69% transforming region of the bovine papilloma virus genome.

14. A mammalian cell in accordance with either one of claim 4 or claim 8, wherein said first and second subunits are from the same mammalian species.

15. A mammalian cell consisting essentially of a cell produced by recombinant DNA techniques, said recombinant cell being capable of producing a biologically active thyroid stimulating hormone (TSH), or progeny of said recombinant cell capable of producing said biologically active heterodimeric protein.

16. A method for producing a biologically active heterodimeric protein comprising culturing the mammalian cells of claim 4 or claim 8.

17. A method for producing a biologically active heterodimeric protein comprising culturing the mammalian cells of claim 5 or claim 9.
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